Shaping the understanding of saliva-derived effectors towards aphid colony proliferation in host plant

‘Effectors’ are proteins and/or small molecules that originated from aphid saliva gland and its secretion is initiated due to interaction between host and insect. The effectors have the ability to manipulate the host cell structure as well as function similar to pathogen’s effectors. Like pathogen’s effectors, aphid effectors suppress the hosts’ defense responses as well as hosts’ defense induction or both. In the susceptible interaction with the host, aphid effectors alter plant processes that contribute to the establishment of compatibility that promotes aphid proliferation. In the susceptible reaction with the host, aphid effectors contribute to the successful salivation and sustainability of the sieve element sap ingestion that have promoting role in more aphid proliferation. In the resistant interaction with the host, aphid effectors are recognized by the typical plant receptors and elicit the induction the effective defense response. As a result, aphid proliferation is reduced due to reduced compatibility establishment in the resistant host. This review focuses on the exciting progress in aphid effector biology that insights new perspective in the molecular basis of plant–aphid interactions.     

The Pseudomonas syringae type III effector HopAM1 enhances virulence on water-stressed plants

Pseudomonas syringae strains deliver diverse type III effector proteins into host cells, where they can act as virulence factors. Although the functions of the majority of type III effectors are unknown, several have been shown to interfere with plant basal defense mechanisms. Type III effectors also could contribute to bacterial virulence by enhancing nutrient uptake and pathogen adaptation to the environment of the host plant. We demonstrate that the type III effector HopAM1 (formerly known as AvrPpiB) enhances the virulence of a weak pathogen in plants that are grown under drought stress. This is the first report of a type III effector that aids pathogen adaptation to water availability in the host plant. Expression of HopAM1 makes transgenic Ws-0 Arabidopsis hypersensitive to abscisic acid (ABA) for stomatal closure and germination arrest. Conditional expression of HopAM1 in Arabidopsis also suppresses basal defenses. ABA responses overlap with defense responses and ABA has been shown to suppress defense against P. syringae pathogens. We propose that HopAM1 aids P. syringae virulence by manipulation of ABA responses that suppress defense responses. In addition, host ABA responses enhanced by type III delivery of HopAM1 protect developing bacterial colonies inside leaves from osmotic stress.     

Entering and breaking: virulence effector proteins of oomycete plant pathogens

Oomycete pathogens of plants and animals are related to marine algae and have evolved mechanisms to avoid or suppress host defences independently of other groups of pathogens, such as bacteria and fungi. They cause many destructive diseases affecting crops, forests and aquaculture. The development of genomic resources has led to a dramatic increase in our knowledge of the effectors used by these pathogens to suppress host defences. In particular, a huge, rapidly diverging superfamily of effectors with 100–600 members per genome has been identified. Proteins in this family use the N-terminal motifs RxLR and dEER to cross the host plasma cell membrane autonomously. Once inside the host cell, the proteins suppress host defence signalling. The importance of this effector family is underlined by the fact that plants have evolved intracellular defence receptors to detect the effectors and trigger a rapid counter-attack. The mechanisms by which the effector enter host cells, and by which they suppress host defences, remain to be elucidated.     

Versatile effectors of phytopathogenic fungi target host immunity

Phytopathogenic fungi secrete a large arsenal of effector molecules, including proteinaceous effectors, small RNAs, phytohormones and derivatives thereof. The pathogenicity of fungal pathogens is primarily determined by these effectors that are secreted into host cells to undermine innate immunity, as well as to facilitate the acquisition of nutrients for their in planta growth and proliferation. After conventional and non-conventional secretion, fungal effectors are translocated into different subcellular compartments of the host cells to interfere with various biological processes. In extracellular spaces, apoplastic effectors cope with physical and chemical barriers to break the first line of plant defenses. Intracellular effectors target essential immune components on the plasma membrane, in the cytosol, including cytosolic organelles, and in the nucleus to suppress host immunity and reprogram host physiology, favoring pathogen colonization. In this review, we comprehensively summarize the recent advances in fungal effector biology, with a focus on the versatile virulence functions of fungal effectors in promoting pathogen infection and colonization. A perspective of future research on fungal effector biology is also discussed.     

Challenges and progress towards understanding the role of effectors in plant-fungal interactions

Both mutualistic and biotrophic pathogenic fungi rely on living host plants for growth and reproduction and must modify host cell structure and function for successful infection. The deployment of a diverse set of secreted virulence determinants referred to as 'effectors', many of which are directly delivered into the host cell, is postulated to be the key to host infection. This review provides a snapshot of the current progress in fungal effector biology. Recent genome sequencing of rust and powdery mildew obligate biotrophs has provided insight into the repertoires of potential effectors of these highly specialised pathogens. Identification of the first host-translocated effectors from mutualistic fungi has revealed that these fungi also manipulate host cells through effectors. The biological activities of some fungal effectors are just beginning to be revealed, while much uncertainty still surrounds the mechanisms of transport into host cells.     

Loss of susceptibility as a novel breeding strategy for durable and broad-spectrum resistance

Recent studies on plant immunity have suggested that a pathogen should suppress induced plant defense in order to infect a plant species, which otherwise would have been a nonhost to the pathogen. For this purpose, pathogens exploit effector molecules to interfere with different layers of plant defense responses. In this review, we summarize the latest findings on plant factors that are activated by pathogen effectors to suppress plant immunity. By looking from a different point of view into host and nonhost resistance, we propose a novel breeding strategy: disabling plant disease susceptibility genes (S-genes) to achieve durable and broad-spectrum resistance.     

Suppression of plant resistance gene-based immunity by a fungal effector

The innate immune system of plants consists of two layers. The first layer, called basal resistance, governs recognition of conserved microbial molecules and fends off most attempted invasions. The second layer is based on Resistance (R) genes that mediate recognition of effectors, proteins secreted by pathogens to suppress or evade basal resistance. Here, we show that a plant-pathogenic fungus secretes an effector that can both trigger and suppress R gene-based immunity. This effector, Avr1, is secreted by the xylem-invading fungus Fusarium oxysporum f.sp. lycopersici (Fol) and triggers disease resistance when the host plant, tomato, carries a matching R gene (I or I-1). At the same time, Avr1 suppresses the protective effect of two other R genes, I-2 and I-3. Based on these observations, we tentatively reconstruct the evolutionary arms race that has taken place between tomato R genes and effectors of Fol. This molecular analysis has revealed a hitherto unpredicted strategy for durable disease control based on resistance gene combinations.     

The zig-zag-zig in oomycete–plant interactions

In addition to a range of preformed barriers, plants defend themselves against microbial invasion by detecting conserved, secreted molecules, called pathogen-associated molecular patterns (PAMPs). PAMP-triggered immunity (PTI) is the first inducible layer of plant defence that microbial pathogens must navigate by the delivery of effector proteins that act to suppress or otherwise manipulate key components of resistance. Effectors may themselves be targeted by a further layer of defence, effector-triggered immunity (ETI), as their presence inside or outside host cells may be detected by resistance proteins. This 'zig-zag-zig' of tightly co-evolving molecular interactions determines the outcome of attempted infection. In this article, we consider the complex molecular interplay between plants and plant pathogenic oomycetes, drawing on recent literature to illustrate what is known about oomycete PAMPs and elicitors of defence responses, the effectors they utilize to suppress PTI, and the phenomenal molecular 'battle' between effector and resistance ( R ) genes that dictates the establishment or evasion of ETI.     

Pathogens drop the hint: don't forget phytoalexin pathways

Many effectors secreted by pathogenic bacteria suppress host signal transduction pathways that activate host defense responses. In this issue of Cell Host & Microbe, Zhou et?al. (2011) now broaden that theme by demonstrating that HopZ1b from Pseudomonas syringae pv. glycinea causes degradation of a host enzyme directly involved in the synthesis of antimicrobial phytoalexin compounds.     

Secretion of type III effectors into host cells in real time

Type III secretion (T3S) systems are key features of many gram-negative bacteria that translocate T3S effector proteins directly into eukaryotic cells. There, T3S effectors exert many effects, such as cellular invasion or modulation of host immune responses. Studying spatiotemporal orchestrated secretion of various effectors has been difficult without disrupting their functions. Here we developed a new approach using Shigella flexneri T3S as a model to investigate bacterial translocation of individual effectors via multidimensional time-lapse microscopy. We demonstrate that direct fluorescent labeling of tetracysteine motif-tagged effectors IpaB and IpaC is possible in situ without loss of function. Studying the T3S kinetics of IpaB and IpaC ejection from individual bacteria, we found that the entire pools of IpaB and IpaC were released concurrently upon host cell contact, and that 50% of each effector was secreted in 240 s. This method allows an unprecedented analysis of the spatiotemporal events during T3S.     

Analysis of Putative Apoplastic Effectors from the Nematode,Globodera rostochiensis,and Identification of an Expansin-Like Protein That Can Induce and Suppress Host Defenses

The potato cyst nematode, Globodera rostochiensis, is an important pest of potato. Like other pathogens, plant parasitic nematodes are presumed to employ effector proteins, secreted into the apoplast as well as the host cytoplasm, to alter plant cellular functions and successfully infect their hosts. We have generated a library of ORFs encoding putative G. rostochiensis putative apoplastic effectors in vectors for expression in planta. These clones were assessed for morphological and developmental effects on plants as well as their ability to induce or suppress plant defenses. Several CLAVATA3/ESR-like proteins induced developmental phenotypes, whereas predicted cell wall-modifying proteins induced necrosis and chlorosis, consistent with roles in cell fate alteration and tissue invasion, respectively. When directed to the apoplast with a signal peptide, two effectors, an ubiquitin extension protein (GrUBCEP12) and an expansin-like protein (GrEXPB2), suppressed defense responses including NB-LRR signaling induced in the cytoplasm. GrEXPB2 also elicited defense response in species- and sequence-specific manner. Our results are consistent with the scenario whereby potato cyst nematodes secrete effectors that modulate host cell fate and metabolism as well as modifying host cell walls. Furthermore, we show a novel role for an apoplastic expansin-like protein in suppressing intra-cellular defense responses.     

Proteases in pathogenesis and plant defence

Plant pathogens deliver a variety of virulence factors to host cells to suppress basal defence responses and create suitable environments for their propagation. Plants have in turn evolved disease resistance genes whose products detect the virulence factors as a signal of invasion and activate effective defence responses. Understanding how a virulence effector contributes to virulence on susceptible hosts but becomes an avirulence factor that triggers defence responses on resistance hosts has been a major focus in plant research. Recent studies have shown that a growing list of pathogen-encoded effectors functions as proteases that are secreted into plant cells to modify host proteins. In addition, several plant proteases have been found to function in activation of the defence mechanism. These findings reveal that post-translational modification of host proteins through proteolytic processing is a widely used mechanism in regulating the plant defence response.     

Terminal reassortment drives the quantum evolution of type III effectors in bacterial pathogens

Many bacterial pathogens employ a type III secretion system to deliver type III secreted effectors (T3SEs) into host cells, where they interact directly with host substrates to modulate defense pathways and promote disease. This interaction creates intense selective pressures on these secreted effectors, necessitating rapid evolution to overcome host surveillance systems and defenses. Using computational and evolutionary approaches, we have identified numerous mosaic and truncated T3SEs among animal and plant pathogens. We propose that these secreted virulence genes have evolved through a shuffling process we have called "terminal reassortment." In terminal reassortment, existing T3SE termini are mobilized within the genome, creating random genetic fusions that result in chimeric genes. Up to 32% of T3SE families in species with relatively large and well-characterized T3SE repertoires show evidence of terminal reassortment, as compared to only 7% of non-T3SE families. Terminal reassortment may permit the near instantaneous evolution of new T3SEs and appears responsible for major modifications to effector activity and function. Because this process plays a more significant role in the evolution of T3SEs than non-effectors, it provides insight into the evolutionary origins of T3SEs and may also help explain the rapid emergence of new infectious agents.     

ExoS controls the cell contact-mediated switch to effector secretion in Pseudomonas aeruginosa

Type III secretion is used by many gram-negative bacterial pathogens to directly deliver protein toxins (effectors) into targeted host cells. In all cases, secretion of effectors is triggered by host cell contact, although the mechanism is unclear. In Pseudomonas aeruginosa, expression of all type III secretion-related genes is up-regulated when secretion is triggered. We were able to visualize this process using a green fluorescent protein reporter system and to use it to monitor the ability of bacteria to trigger effector secretion on cell contact. Surprisingly, the action of one of the major type III secreted effectors, ExoS, prevented triggering of type III secretion by bacteria that subsequently attached to cells, suggesting that triggering of secretion is feedback regulated. Evidence is presented that translocation (secretion of effectors across the host cell plasma membrane) of ExoS is indeed self-regulated and that this inhibition of translocation can be achieved by either of its two enzymatic activities. The translocator proteins PopB, PopD, and PcrV are secreted via the type III secretion system and are required for pore formation and translocation of effectors across the host cell plasma membrane. Here we present data that secretion of translocators is in fact not controlled by calcium, implying that triggering of effector secretion on cell contact represents a switch in secretion specificity, rather than a triggering of secretion per se. The requirement for a host cell cofactor to control effector secretion may help explain the recently observed phenomenon of target cell specificity in both the Yersinia and P. aeruginosa type III secretion systems.     

Coiled-coil proteins associated with type III secretion systems: a versatile domain revisited

The pathogenic potential of many Gram-negative bacteria is indicated by the possession of a specialized type III secretion system that is used to deliver virulence effector proteins directly into the cellular environment of the eukaryotic host. Extracellular assemblies of secreted proteins contrive a physical link between the pathogen and host cytosol and enable the translocated effectors to bypass the bacterial and host membranes in a single step. Subsequent interactions of some effector proteins with host cytoskeletal and signalling proteins result in modulation of the cytoskeletal architecture of the aggressed cell and facilitate entry, survival and dissemination of the pathogen. Although the secreted components of type III secretion systems are diverse, many are predicted to share a common coiled-coil structural feature. Coiled-coils are ubiquitous and highly versatile assembly motifs found in a wide range of structural and regulatory proteins. The prevalence of these domains in secreted virulence effector proteins suggests a fundamental contribution to multiple aspects of their function, and evidence accumulating from functional studies suggests an intrinsic involvement of coiled-coils in subunit assembly, translocation and flexible interactions with multiple bacterial and host proteins. The known functional flexibility that coiled-coil domains confer upon proteins provides insights into some of the pathogenic mechanisms used during interaction with the host.     

Common and contrasting themes in host cell-targeted effectors from bacterial,fungal, oomycete and nematode plant symbionts described using the Gene Ontology

A wide diversity of plant-associated symbionts, including microbes, produce proteins that can enter host cells, or are injected into host cells in order to modify the physiology of the host to promote colonization. These molecules, termed effectors, commonly target the host defense signaling pathways in order to suppress the defense response. Others target the gene expression machinery or trigger specific modifications to host morphology or physiology that promote the nutrition and proliferation of the symbiont. When recognized by the host's surveillance machinery, which includes cognate resistance (R) gene products, defense responses are engaged to restrict pathogen proliferation. Effectors from diverse symbionts may be delivered into plant cells via varied mechanisms, including whole organism cellular entry (viruses, some bacteria and fungi), type III and IV secretion (in bacteria), physical injection (nematodes and insects) and protein translocation signal sequences (oomycetes and fungi). This mini-review will summarize both similarities and differences in effectors and effector delivery systems found in diverse plant-associated symbionts as well as how these are described with Plant-Associated Microbe Gene Ontology (PAMGO) terms.     

Oomycete and fungal effector entry, a microbial Trojan horse

Oomycete and fungal symbionts have significant impacts on most commercially important crop and forest species, and on natural ecosystems, both negatively as pathogens and positively as mutualists. Symbiosis may be facilitated through the secretion of effector proteins, some of which modulate a variety of host defense mechanisms. A subset of these secreted proteins are able to translocate into host cells. In the oomycete pathogens, two conserved N-terminal motifs, RXLR and dEER, mediate translocation of effector proteins into host cells independent of any pathogen-encoded machinery. An expanded 'RXLR-like' motif [R/K/H]X[L/M/I/F/Y/W]X has been used to identify functional translocation motifs in host-cell-entering fungal effector proteins from pathogens and a mutualist. The RXLR-like translocation motifs were required for the fungal effectors to enter host cells in the absence of any pathogen-encoded machinery. Oomycete and fungal effectors with RXLR and RXLR-like motifs can bind phospholipids, specifically phosphatidylinositol-3-phosphate (PtdIns-3-P). Effector-PtdIns-3-P binding appears to mediate cell entry via lipid raft-mediated endocytosis, and could be blocked by sequestering cell surface PtdIns-3-P or by utilizing inositides that competitively inhibit effector binding to PtdIns-3-P. These findings suggest that effector blocking technologies could be developed and utilized in a variety of important crop species against a broad spectrum of plant pathogens.     

Comparative Large-Scale Analysis of Interactions between Several Crop Species and the Effector Repertoires from Multiple Pathovars of Pseudomonas and Ralstonia

Bacterial plant pathogens manipulate their hosts by injection of numerous effector proteins into host cells via type III secretion systems. Recognition of these effectors by the host plant leads to the induction of a defense reaction that often culminates in a hypersensitive response manifested as cell death. Genes encoding effector proteins can be exchanged between different strains of bacteria via horizontal transfer, and often individual strains are capable of infecting multiple hosts. Host plant species express diverse repertoires of resistance proteins that mediate direct or indirect recognition of bacterial effectors. As a result, plants and their bacterial pathogens should be considered as two extensive coevolving groups rather than as individual host species coevolving with single pathovars. To dissect the complexity of this coevolution, we cloned 171 effector-encoding genes from several pathovars of Pseudomonas and Ralstonia. We used Agrobacterium tumefaciens-mediated transient assays to test the ability of each effector to induce a necrotic phenotype on 59 plant genotypes belonging to four plant families, including numerous diverse accessions of lettuce (Lactuca sativa) and tomato (Solanum lycopersicum). Known defense-inducing effectors (avirulence factors) and their homologs commonly induced extensive necrosis in many different plant species. Nonhost species reacted to multiple effector proteins from an individual pathovar more frequently and more intensely than host species. Both homologous and sequence-unrelated effectors could elicit necrosis in a similar spectrum of plants, suggesting common effector targets or targeting of the same pathways in the plant cell.